Exosome-Enriched Plasma Analysis as a Tool for the Early Detection of Hypertensive Gestations

Abstract

Hypertensive disorders of pregnancy are closely associated with prematurity, stillbirth, and maternal morbidity and mortality. The onset of hypertensive disorders of pregnancy (HDP) is generally noticed after the 20th week of gestation, limiting earlier intervention. The placenta is directly responsible for modulating local and systemic physiology by communicating using mechanisms such as the release of extracellular vesicles, especially exosomes. In this study, we postulated that an analysis of exosome-enriched maternal plasma could provide a more focused and applicable approach for diagnosing HDP earlier in pregnancy. Therefore, the peripheral blood plasma of 24 pregnant women (11 controls, 13 HDP) was collected between 20th and 24th gestational weeks and centrifuged for exosome enrichment. Exosome-enriched plasma samples were analyzed by Raman spectroscopy and by proton nuclear magnetic resonance metabolomics (1H NMR). Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to analyze the Raman data, from the spectral region of 600–1,800 cm–1, to determine its potential to discriminate between groups. Using principal component analysis, we were able to differentiate the two groups, with 89% of all variances found in the first three principal components. In patients with HDP, most significant differences in Raman bands intensity were found for sphingomyelin, acetyl CoA, methionine, DNA, RNA, phenylalanine, tryptophan, carotenoids, tyrosine, arginine, leucine, amide I and III, and phospholipids. The 1H NMR analysis showed reduced levels of D-glucose, L-proline, L-tyrosine, glycine, and anserine in HDP, while levels of 2-hydroxyvalerate, polyunsaturated fatty acids, and very-low-density lipoprotein (VLDL) were increased. 1H NMR results were able to assign an unknown sample to either the control or HDP groups at a precision of 88.3% using orthogonal partial least squares discriminant analysis and 87% using logistic regression analysis. Our results suggested that an analysis of exosome-enriched plasma could provide an initial assessment of placental function at the maternal-fetal interface and aid HDP diagnosis, prognosis, and treatment, as well as to detect novel, early biomarkers for HDP.

Circulating extracellular vesicles in healthy and pathological pregnancies: A scoping review of methodology, rigour and results

“…One study did not include a healthy pregnant control group (Abolbaghaei et al., 2021). Pathologies included were: pre‐eclampsia (PE; n = 27) (Alasztics et al., 2021; Alijotas et al., 2012; Campello et al., 2015; Dragovic et al., 2013; Hu et al., 2018; Jadli et al., 2017; Kovács et al., 2018a, 2018b; Lalic‐Cosic et al., 2021; León et al., 2021; Li et al., 2020; Lok et al., 2007, 2008, 2009b; Maduray et al., 2020; Marques et al., 2012; Mikhailova et al., 2014; Murugesan et al., 2022; Pillay et al., 2016, 2020; Salomon et al., 2017; Textoris et al., 2013; Truong et al., 2017; VanWijk et al., 2002a; Verma et al., 2018; Wang et al., 2020; Weingrill et al., 2021; Zhang et al., 2018), small for gestational age/fetal growth restriction/intrauterine growth restriction (SGA/FGR/IUGR; n = 5) (Alijotas et al., 2012; Bretelle et al., 2003; Jadli et al., 2017; Li et al., 2020; Miranda et al., 2018), pre‐term birth or premature pre‐term rupture of membranes (PTB/PPROM; n = 4) (Menon et al., 2020; Menon, Debnath et al., 2019; Tronco et al., 2020; Truong et al., 2017), PE with IUGR ( n = 2) (Alijotas et al., 2012; Jadli et al., 2017), gestational diabetes mellitus (GDM; n = 4) (James‐Allan et al., 2020; Jiang et al., 2022; Salomon et al., 2016; Zhang et al., 2021), pregnancy loss/recurrent miscarriage/unexplained fetal loss (PL/RM/UFL; n = 2) (Alijotas‐Reig et al., 2011; Monteiro et al., 2020), Type‐1 diabetes ( n = 1) (Abolbaghaei et al., 2021), HIV with or without PE ( n = 1) (Pillay et al., 2020), HIV with or without malaria ( n = 1) (Moro et al.,…”

“…One study did not include a healthy pregnant control group (Abolbaghaei et al, 2021). Pathologies included were: pre-eclampsia (PE; n = 27) (Alasztics et al, 2021;Alijotas et al, 2012;Campello et al, 2015;Dragovic et al, 2013;Hu et al, 2018;Jadli et al, 2017;Kovács et al, 2018aKovács et al, , 2018bLalic-Cosic et al, 2021;León et al, 2021;Li et al, 2020;Lok et al, 2007Lok et al, , 2008Lok et al, , 2009bMaduray et al, 2020;Marques et al, 2012;Mikhailova et al, 2014;Murugesan et al, 2022;Pillay et al, 2016Pillay et al, , 2020Textoris et al, 2013;Truong et al, 2017;VanWijk et al, 2002a;Verma et al, 2018;Wang et al, 2020;Weingrill et al, 2021;Zhang et al, 2018), small for gestational age/fetal growth restriction/intrauterine growth restriction (SGA/FGR/IUGR; n = 5) (Alijotas et al, 2012;Bretelle et al, 2003;Jadli et al, 2017;Li et al, 2020;Miranda et al, 2018), pre-term birth or premature preterm rupture of membranes (PTB/PPROM; n = 4) (Menon et al, 2020;Menon, Debnath et al, 2019;Tronco et al, 2020;Truong et al, 2017) (Pillay et al, 2020), HIV with or without malaria (n = 1) (Moro et al, 2016) and anti-phospholipid syndrome (APS; n = 1) (Campello et a...…”

Metabolomics of Extracellular Vesicles: A Future Promise of Multiple Clinical Applications

“… 68 Furthermore, Weingrill et al found that plasma EVs could also be used to provide an initial assessment of placental function and to develop diagnosis, treatment, and prognosis of gestational hypertension. 88 Notably, for ST elevation myocardial infarction, evidence has demonstrated that plasma EVs in patients contain more lipid molecules, revealing possible biomarkers and signal transduction mechanisms. 69 …”

“… Diagnosis Late myocardial necrosis of atherosclerotic Adipose tissue C16:0 ceramide and their derivatives [ 87 ] Acute pancreatitis Plasma Eicosatrienoic acid (C20:3), thiamine triphosphate. 2-Acetylfuran, cis-Citral [ 68 ] Gestational hypertension Plasma SM, acetyl CoA, methionine, phenylalanine, tryptophan, carotenoids, tyrosine, arginine, leucine [ 88 ] ST elevation myocardial infarction Plasma Ceramides, dihydroceramides, and sphingomyelins [ 69 ] Lung cancer Pleural effusions Phenylalanine, leucine, PC 35:0 and SM 44:3 [ 91 ] Serum Ceramides, PC and polyunsaturated acyl chains [ 92 ] Prostate cancer Urinary PS 18:1/18:1 and lactosylceramide (d18:1/16:0) [ 93 ] Benign prostate hyperplasia Urinary PC, PE, SM [ 31 ] Bipolar disorder Serum lysoPE 18: 0, lysoPE 14: 0, lysoPC 18: 0, and lysoPC 20: 1 [ 73 ] Schizophrenia Depressive disorder Chromochromic leukogystrophy …”

Exosomes in the Treatment of Pancreatic Cancer: A Moonshot to PDAC Treatment?

“…It is now common knowledge that they are important mediators of intercellular communication under both normal and diseased conditions [ 49 ]. In more detail, the mediation of exosomes regulates a wide variety of physiologic processes: skin regeneration [ 50 ], bone remodeling [ 51 ], placental formation [ 52 ], and organ crosstalk during exercise [ 53 ]. It also serves as a diagnostic and predictive biomarker in inflammatory diseases such as rheumatoid arthritis [ 54 ], systemic lupus erythematosus [ 55 ], and nephropathies [ 56 ].…”